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Exosome and preparing method and application thereof

A technology of exosomes and in vitro culture, applied in the direction of pharmaceutical formulations, medical preparations containing active ingredients, bone/connective tissue cells, etc., can solve problems such as cognitive dysfunction

Active Publication Date: 2019-05-21
INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Aiming at how to solve the problem of postoperative cognitive dysfunction, the present invention provides an exosome, which is antler mesenchymal stem cell-derived exosome

Method used

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  • Exosome and preparing method and application thereof
  • Exosome and preparing method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1. Preparation method of exosomes derived from velvet antler mesenchymal stem cells.

[0054] 1. Primary isolation and culture of velvet antler mesenchymal stem cells: After pretreatment of velvet antler tissue, the mesenchymal area was obtained, and mesenchymal stem cells were isolated. Methods as below:

[0055] Pretreatment: Collect fresh antler in late spring and early summer. First, squeeze the blood out of the sawn antler, then wrap it in tin foil, then wrap it in cotton, and put it in a foam box with an ice box. Cut the antler tip tissue about 5 cm long, then squeeze the blood, then cut into 4 halves (longitudinal), then wash 3 times with 75% alcohol for 2 minutes each time, and finally wash 3 times with PBS. Then, put the processed velvet antler into the ultra-clean bench to peel off the dermis, then wash it 3 times with PBS, wash it with 10% fetal bovine serum and double antibody (final concentration is 100U / mL penicillin and final concentration is 10...

Embodiment 2

[0070] Example 2. The role of antler mesenchymal stem cells (AMSCs) exosomes in the treatment of cognitive dysfunction.

[0071] Taking mice as the experimental subjects, the effect of the antler mesenchymal stem cells (AMSCs) exosomes prepared in Example 1 in the treatment of cognitive dysfunction was investigated.

[0072] 1. Experimental animals and grouping:

[0073] Forty male SD rats of SPF grade, weighing 350-450g, were randomly divided into 4 groups, 10 rats in each group, namely: sham operation group (Sham group), CPB operation group (CPB group), exosome + CPB group (EXO group) and exosomes + TLR2 / TLR4 inhibitor group (TLR group), except for the Sham group, were all prepared by CPB bypass to create a CPB rat POCD model.

[0074] Preparation of CPB model:

[0075] Before the operation, the experimental animals were fasted for 6 hours, anesthetized by intraperitoneal injection of pentobarbital sodium 30mg / kg, fixed on the operating table after the righting reflex disa...

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Abstract

The invention discloses an exosome and a preparing method and application thereof, and relates to the field of exosome medicines. In order to solve the problem of postoperative cognition dysfunction,mesenchymal stem cells are separated out of cornu cervi tissue; after culturing, a cell culture solution is collected, and the exosome from cornu cervi mesenchymal stem cells are extracted through separation. It is proved that CPB rat nerve function deficit and oxidative stress response can be relived, a protection effect is realized for CPB rat nerve cell apoptosis and CPB rat POCD can be relieved through a TLR2 / TLR4 signal channel. The exosome can be used for preparing medicines for improving or treating the cognition dysfunction.

Description

technical field [0001] The invention relates to the field of exosome medicine, in particular to an exosome and its preparation method and application. Background technique [0002] With the rapid development of cardiopulmonary bypass (CPB) technology and anesthesia technology, the mortality rate caused by complications of the central nervous system has increased. Postoperative cognitive dysfunction (POCD) is the Severe complications of surgery, and the inflammatory response induced by CPB is the main factor that promotes POCD. Therefore, how to prevent and treat POCD induced by CPB cardiac surgery has become a problem that plagues the development of cardiac surgery. [0003] Mesenchymal stem cells currently used in the treatment of diseases are all derived from different human tissues, such as bone marrow, umbilical cord and fat. However, current studies have shown that these three types of cells are not specific in the treatment of senile neurodegenerative diseases. In a...

Claims

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Application Information

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IPC IPC(8): C12N5/0775A61K35/28A61P25/28
Inventor 杨春孙胜楠彭英华张琦郭佳杨敏刘莹
Owner INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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